Effects of storage duration and storage media on initial and post-eyeing mortality of stored ova of rainbow trout Oncorhynchus mykiss

The effects of storage duration and storage media on the viability of unfertilized eggs of rainbow trout from fertilization to hatching stage was studied. The unfertilized eggs were stored in coelomic fluid and Cortland artificial media buffered with Tris–HCl (C₄H₁₁NO₃–HCl) and Hepes (C₈H₁₈N₂O₄S) at 2–3 °C. These eggs were fertilized after 0 (i.e. control eggs fertilized prior to storage), 2 and 9 days post-stripping using fresh, pooled sperm obtained from 4 to 5 males. According to the results of present study, after 2 days of storage, no significant (p > 0.05) change in survival to eyeing and hatching of eggs stored in coelomic fluid (85.5 ± 4.8%, 80.2 ± 6.3%) and Cortland medium buffered with Hepes (73.3 ± 4.1%,68.1 ± 4%) was observed in comparison to the control (82.6 ± 8%,78.7 ± 7.8%). However, eyeing and hatching survival rates of eggs stored for the same period in Cortland medium buffered with Tris–HCl (44.5 ± 7%,34.2 ± 8.2%) decreased significantly (p < 0.05) in comparison with the control. Also, eggs stored for 2 days in Cortland medium buffered with Tris–HCl (25 ± 6.7%) had significantly (p < 0.05) greater eyed egg mortality compared to the control (4.7 ± 0.6%), while no significant (p > 0.05) differences were observed between eyed egg mortality rates of eggs stored in coelomic fluid (6.7 ± 2.1%) and Cortland medium buffered with Hepes (7.3 ± 0.9%) compared to control. After 9 days of storage, the eggs that were stored in coelomic fluid showed no significant (p > 0.05) differences in eyeing (77.5 ± 7.2%) and hatching (67 ± 8.1%) rates in comparison with control, while both Cortland storage media exhibited significant (p < 0.05) loss in eyeing (44.4 ± 11.5%, 24.7 ± 13.8% for Hepes and Tris–HCl, respectively), and hatching (24.5 ± 5.9%, 19.2 ± 14.4% for Hepes and Tris–HCl, respectively) in comparison with control. Furthermore, eyed egg mortality increased significantly (p < 0.05) in coelomic fluid (14 ± 2.9%), media buffered with Hepes (43 ± 6.2%) and Tris–HCl (49.3 ± 26.9%) compared to control. Based on this study, unfertilized eggs of rainbow trout can be successfully stored in coelomic fluid for 9 days at (2–3 °C) without significant loss of fertility. However, storage within a similar period in artificial media showed lower fertilization rates and higher eyed egg mortality compared to coelomic fluid.     

Studies on cryopreservation of eggs from rainbow trout (Salmo gairdneri) and coho salmon (Oncorhynchus kisutch)

The effect of pre-freezing treatments as well as freezing of inseminated, not water-activated eggs from rainbow trout Salmo gairdneri, and coho salmon, Oncorhynchus kisutch, was investigated in relation to survival and further development.Effects above freezing temperatures included: the temperature at insemination, viability of inseminated and unactivated eggs after storage, suitability of an incubation medium and the tolerance of eggs to various levels of the cryoprotectant dimethylsulfoxide (DMSO). Freezing experiments included: investigating the action of DMSO (0, 1, 2 mole) and the tolerance of coho eggs to temperatures between ?4.6 to ?30°C. Insemination temperatures between 0.5°C and 9.8°C (coho eggs) as well as incubation in an artifical medium (1-0°C) for 80 min (rainbow trout eggs) and 170 min (coho eggs) did not influence subsequent fertility. Storage of inseminated and unactivated rainbow trout eggs for 135 min and beyond reduced egg fertility. DMSO at 2 and 4 mole was detrimental to coho eggs (1-0°C). One mole DMSO had no (coho) or reduced (rainbow trout) influence on egg fertility when it was added gradually.In the presence of 1 mole DMSO most eggs remained unfrozen (67–89%) when kept for 10 min in frozen artificial medium (?4.6%) and 27–32% subsequently reached the eyed stage (control = 100). Further cooling (0.3°C/min) to ?10°C was still tolerated (62% unfrozen, 22% eyed eggs) but not to ?20°C (6% unfrozen, no development) and ?30°C (no survival). Use of 2 mole DMSO did not improve the results.     

Preservation of black sharkminnow,Labeo chrysophekadion (Bleeker, 1849) spermatozoa

The objective of this study was to investigate the effects of extenders and storage time on motility, viability and fertilization of preserved black sharkminnow, Labeo chrysophekadion spermatozoa. Sperm were diluted 1:3 in one of five extenders: modified Cortland solution (MC); Hanks' balanced salt solution (HBSS); 0.9% sodium chloride (NaCl); Kurokura solution (KU); and modified extender, and undiluted sperm samples were used as control and stored at 4°C for 5 days. Motility, viability and fertilization rates were evaluated every day. After a storage time of three days, the highest motility, viability and fertilization rates (61.27 ± 2.26%, 58.60 ± 2.29% and 40.58 ± 0.57, respectively) were achieved with sperm diluted with modified extender. Motility, viability and fertilization rates decreased significantly (P < 0.05) with increasing storage time in all treatments. In addition, this study found that motility, viability and fertilization had a positive significant correlation (P < 0.01). The results indicate that isotonic extender is suitable for the short‐term preservation of black sharkminnow spermatozoa.     

Determination of growth performance,meat quality and colour attributes of large rainbow trout (Oncorhynchus mykiss) in the southern Black Sea coasts of Turkey

The aim of this study was to determine the growth performance, meat quality and colour attributes of large rainbow trout (Oncorhynchus mykiss) in net cage systems in the southern Black Sea coasts of Turkey. In the study, large rainbow trout with an initial weight of 1,322.07 ± 57.72 g were reached to 3,385.30 ± 140.98 g and the survival rates were 95.69 ± 1.32% in 5‐month production period. Crude protein and crude fat values of fish meat were ranged between 16.75 ± 0.07–20.18 ± 0.90% and 9.22 ± 0.15–12.29 ± 0.42%, respectively (p < .05). The essential and non‐essential amino acid values were between 12.92 ± 0.02–13.09 ± 0.01 g/100 g and 11.97 ± 0.14–11.47 ± 0.01 g/100 g, respectively (p > .05). C20:5n‐3, C22:6n‐3, ?3 and ?6 values in fish meat were between 4.32 ± 0.02–4.30 ± 0.08% (p < .05), 8.31 ± 0.10–7.72 ± 0.15% (p < .05), 13.41 ± 0.10–12.93 ± 0.23% (p < .05) and 21.85 ± 0.06%‐19.74 ± 0.36 (p < .05), respectively. As a result, it can be concluded that large rainbow trout cultured in the southern Black Sea coasts of Turkey is a rich source of food in terms of fatty and amino acid compositions and atherogenicity index, thrombogenicity index and hypocholesterolaemic/hypercholesterolaemic values.     

Egg hatching rate and fatty acid composition of Acartia bilobata (Calanoida,Copepoda) across cold storage durations

To investigate egg storage capacity of the copepod Acartia bilobata for aquaculture interest, we tested hatching success rate (HSR) of inclusive eggs (mixture of all egg types) after 4°C storage. The HSR peaked after 14 days storage when incubating at 28°C for 48 hr (85.8 ± 1.6%) and 72 hr (87.6 ± 0.9%), then gradually declined until 1 year (48 hr: 7 ± 0.6%; 72 hr: 19.4 ± 3.9%). Reallocation of fatty acid profile suggests that docosahexaenoic acid (DHA) is correlated with the HSR of A. bilobata eggs. Additionally, we investigated the HSR of diapausing eggs (unhatched eggs after 72 hr incubation of the inclusive eggs) after 4°C storage. Their HSR peaked after 14 days storage (48 hr:75.3 ± 3.5%; 72 hr:78.2 ± 2.1%), then gradually declined until 60 days (48 hr HSR:42.1 ± 2.3%; 72 hr HSR:53.0 ± 3.2%). Overall, we illustrated the hatchability of diapausing and quiescent eggs of A. bilobata after 4°C storage. The cold storage capacities were low (<60% HSR after 60 days), and it could be limited by the egg DHA content. Our findings provide implications for future studies aiming to improve cold storage techniques of tropical copepod eggs for aquaculture applications.     

Effects of rearing density on growth,fatty acid profile and bioremediation ability of polychaete Nereis diversicolor in an integrated aquaculture system with rainbow trout (Oncorhynchus mykiss)

The objective of this study was to assess the effects of rearing density on the growth and fatty acid profile of Nereis diversicolor and on its capability to bioremediate wastewater in an integrated culture system with rainbow trout (Oncorhynchus mykiss). To this end, a batch of juvenile N. diversicolor (0.03 ± 0.01 mg) was assigned into four different densities (T₁ = 250, T₂ = 500, T₃ = 1,000, T₄ = 2,000, besides T₀ = with no worm) in three replicates. The worm groups were fed with solid waste that was supplied from tanks containing rainbow trout (107.17 ± 13.5 g; 1.39 ± 0.18 kg/m³). During the experiment (60 days), the water temperature was 17.71 ± 0.6°C. The results revealed that SR%, SGR% and WG% of N. diversicolor in T₁ were significantly higher than those of T₂, T₃ and T₄. Both FER rate and biomass gain in T₄ were significantly higher than those in the other groups. The highest removal rate of NO₂‐N (73.72%), NH₃‐N (65.70%), PO₄‐P (59.32%), BOD₅ (69.60%) and TSS (82.33%) were observed in T₄. The major fatty acids presents were palmitic acid, oleic acid, linoleic acid, stearic acid and alpha‐linolenic acid in all worm‐treated groups, with no difference observed in the concentration of these fatty acids among them. Taken together, these findings suggest that organic wastes from rainbow trout farms could be recycled to achieve a sustainable aquaculture goal, and demonstrate that a high percentage of fatty acids in fish feed is not absorbed by rainbow trout but is done by N. diversicolor.     

Dietary supplementation of probiotics and influence on feed efficiency,growth parameters and reproductive performance in female rainbow trout (Oncorhynchus mykiss) broodstock

The aim of this study was to determine the effects of dietary probiotics on reproductive performance, growth, haematological and biochemical parameters of female rainbow trout broodstock. A total of 60 broodstocks with an average initial weight of 2,267.4 ± 54.9 g were divided into four groups and three replicates in raceway ponds. Fish were fed with diets containing 0 (control), 1 × 10⁹ (P1), 2 × 10⁹ (P2) and 4 × 10⁹ (P3) CFU probiotic per kg diet twice a day for 8 weeks before spawning season. Results revealed that higher levels of dietary probiotics (P3) enhanced protein efficiency ratio and decreased feed conversion ratio (p < .05). There were no significant differences in haematological and other growth parameters among different treatments. According to the results, probiotic‐fed treatments had the lowest levels of cholesterol (237.7 ± 4.3 mg/dl) and the highest levels of total protein (11.3 ± 0.3 g/dl) and albumin (6.9 ± 0.3 g/dl), so that differences were found between P3 and control groups in all cases (p < .05). Based on the observations from reproductive parameters, the highest egg diameter, working and relative fecundities, fertilization and hatching rates, eyed eggs survival and alevins survival rates up to the absorption of the yolk sac were observed in P3 treatment (p < .05). Moreover, eyeing, hatching and yolk sac absorbing stages in P3 treatment were earlier than those in other treatments (p < .05). The results of present study demonstrated that probiotic supplementation improved reproductive performance and some growth and biochemical parameters in female rainbow trout broodstock and the best performance was observed in fish fed 4 × 10⁹ CFU probiotic per kg diet. Due to an increase in egg production and egg size in probiotic‐fed treatments, this feed additive can be used as a suitable ingredient to increase high‐quality egg production in rainbow trout breeders.     

Effects of oral passive immunization against somatostatin‐14 on growth performance,body composition and IgY delivery in rainbow trout (Oncorhynchus mykiss) and common carp (Cyprinus carpio)

Egg‐yolk antibodies (IgY) against somatostatin‐14 (anti‐SST‐14) were evaluated as orally administered, growth promotants in gastric rainbow trout (Oncorhynchus mykiss) and agastric common carp (Cyprinus carpio) in an 8‐week feeding trial. Feeding groups were compared with fish which did not receive anti‐SST‐14 IgY. Growth responses and IgY plasma contents of the blood were assessed. In contrast to rainbow trout, oral anti‐SST‐14 addition to carp significantly improved protein efficiency ratio (PER, 1.02 ± 0.04) and protein productive value (PPV, 26.7 ± 0.91) after 56 days (P < 0.05) compared to the negative control (PER = 0.91 ± 0.02; PPV = 22.9 ± 0.66). IgY was undetectable in plasma of rainbow trout after oral administration, indicating that gastric degradation of the fed IgY makes this application route for growth promotion in gastric trout challenging.     

Enhanced growth and retarded gonadal development of farmed rainbow trout,Oncorhynchus mykiss (Walbaum) following a long‐day photoperiod

Long‐day photoperiods are considered as an effective managerial tool in manipulating of reproduction and somatic growth in a number of fish species. In this study, the effects of three different artificial photoperiods on the gonadal development and somatic growth of rainbow trout (Oncorhynchus mykiss L.) were investigated. Two years‐old immature female rainbow trout (279.94 ±2.25 g) were exposed to three artificial photoperiod regimes of 24L:0D, 18L:6D and 6L:18D and natural light (NL) regime for 5 months. The highest gonadosomatic indices were recorded in NL and 6L:18D groups while the rates were significantly lower in fish maintained under 18L:6D and 24L:0D photoperiods (P < 0.05). Mean oocyte diameters in fish exposed to 24L:0D and also to 18L:6D were significantly lower than the 6L:18D and NL groups. Photoperiods with 24L:0D and 18L:6D regimes resulted in significantly higher mean final weights and specific growth rates (SGR) than NL regime. The highest mean final weight (635.45 ± 16.19 g) and SGR (1.03 ± 0.04% day^?1) were obtained under 24L:0D photoperiod. Fish exposed to 24L:0D and 18L:6D showed the highest condition factor as 1.44 ± 0.01 and 1.44 ± 0.02 respectively, when compared with the NL (1.27 ± 0.01) and 6L:18D (1.34 ± 0.02) groups. Basically, the results suggested that continuous artificial lightning can be used as an influential factor in delaying gonadal development and enhancing somatic growth in rainbow trout during gonadal growth phase.     

An investigation on the gamete quality of Black Sea trout (Salmo trutta labrax) broodstock fed with mealworm (Tenebrio molitor)

In this study, the effect of combined feeding Black Sea trout (Salmo trutta labrax) broodstock with commercial feed and mealworm larvae (Tenebrio molitor) for 50 days before the breeding period in order to evaluate the gamete quality. While the control group was fed with commercial feed, mealworm larvae were given as an additional protein source 2 days a week and 3 days a week to the experimental groups formed from female and male individuals. In addition to the growth parameters, the number of eggs (number/individual) and the egg diameter (mm), sperm volume (ml), density of spermatozoa (×10⁹ cell/ml), total motility (%), progressive motility (%) and average curvilinear velocity (VCL, µm/sec) values were determined. At the end of the study, the highest live weight gain was found as 46.2 g in the control group of male. The egg diameters were 4.3 ± 1.8 mm and 4.5 ± 1.4 mm in the worm treatment groups fed twice and three times with mealworm respectively. As a result, it was determined that feeding fish with mealworm larvae as an addition to the commercial diet in female individuals did not affect the amount of eggs (p > .05), but the egg diameters were significantly smaller in the control group than the mealworm groups (p < .05). In spermatological characteristics, only the amount of sperm was different between the groups (p < .05); all other parameters were found to be similar to each other with no statistical differences (p > .05).     

Composition of ovarian fluid in endangered Caspian brown trout, <Emphasis Type="Italic">Salmo trutta caspius</Emphasis>, and its effects on spermatozoa motility and fertilizing ability compared to freshwater and a saline medium

In this study, ovarian fluid composition and its effects on the motility and fertilizing ability of sperm were studied in endangered Caspian brown trout, Salmo trutta caspius, and were compared with a saline activation medium (125 mM NaCl, 30 mM Glycine, 20 mM Tris–HCl, pH = 9.0) and freshwater as the control. The ovarian fluid was composed of sodium 164.4 ± 4.4 mM l⁻¹, potassium 1.8 ± 0.1 mM l⁻¹, calcium 0.6 ± 0.1 mM l⁻¹, magnesium 0.4 ± 0.02 mM l⁻¹, chloride 127.4 ± 5.9 mM l⁻¹, total protein 389.5 ± 89.6 mg 100 ml⁻¹, cholesterol 9.3 ± 1.2 mg dl⁻¹, and glucose 3.3 ± 0.2 mM l⁻¹. The percentage of motile spermatozoa and the duration of sperm motility were significantly higher in ovarian fluid (62 ± 3%, 74.6 ± 0.8 s) than freshwater (35 ± 4%, 44 ± 1 s), but they did not differ significantly from saline medium (56 ± 3%, 74.3 ± 0.7 s) (P > 0.05). Higher eyeing rates were observed after the activation of sperm in ovarian fluid and saline solution than freshwater when 35,000 or 350,000 spermatozoa per egg were added into the activation media. However, no significant differences were observed at higher concentrations of spermatozoa per egg (730,000) (P > 0.05). Also, this study showed that the ovarian fluid composition can be considered as a species-specific character among salmonid fishes. As a conclusion, the results of this study recommend the use of ovarian fluid or the saline solution as an activation medium in the artificial reproduction of Caspian brown trout.     

Purified deoxynivalenol or feed restriction reduces mortality in rainbow trout,Oncorhynchus mykiss (Walbaum), with experimental bacterial coldwater disease but biologically relevant concentrations of deoxynivalenol do not impair the growth of Flavobacterium psychrophilum

Diets containing deoxynivalenol (DON) were fed to rainbow trout Oncorhynchus mykiss (Walbaum) for 4 weeks followed by experimental infection (intraperitoneal) with Flavobacterium psychrophilum (4.1 × 10⁶ colony‐forming units [CFU] mL⁻¹). Mortality of rainbow trout fed either 6.4 mg kg⁻¹ DON or trout pair‐fed the control diet was significantly reduced (P < 0.05) in comparison with trout fed the control diet to apparent satiation (<0.1 mg kg⁻¹ DON). In a second experiment, trout were fed one of three experimental diets; a control diet, a diet produced with corn naturally contaminated with DON (3.3 mg kg⁻¹ DON) or a diet containing purified DON (3.8 mg kg⁻¹); however, these fish were not experimentally infected. The presence of DON resulted in significant reduction (P < 0.0001) in feed intake as well as weight gain after 4 weeks. Respiratory burst of head‐kidney leucocytes isolated from rainbow trout fed diets containing purified DON (3.8 mg kg⁻¹) was significantly higher (P < 0.05) at 35 day post‐exposure compared with controls. The antimicrobial activity of DON was examined by subjecting F. psychrophilum in vitro to serial dilutions of the chemical. Complete inhibition occurred at a concentration of 75 mg L⁻¹ DON, but no effect was observed below this concentration (0–30 mg L⁻¹).     

Biocontrol of saprolegniosis in rainbow trout (Oncorhynchus mykiss Walbaum) using two bacterial isolates (LE89 and LE141) of Pseudomonas fluorescens

The probiotic activity of 15 bacterial isolates that inhibit Saprolegnia parasitica in vitro was tested for the biocontrol of saprolegniosis in rainbow trout (Oncorhynchus mykiss Walbaum), adding the bacteria to tank water for 14 days at a concentration of 10⁶ bacteria ml^?1 water. Pseudomonas fluorescens LE89 and Pseudomonas fluorescens LE141 were effective in controlling experimental infection with S. parasitica since of the fish treated with LE89, 24.5% ± 16.27% (p < 0.05) became infected, as did 42.8% ± 8.41% (p < 0.05) of those treated with LE141. Given their protective effect when administered in water, their effect was also studied when administered in feed before and after experimental infection. Both bacterial isolates survived low pH levels and the action of bile, grew in skin and intestinal mucus, were resistant to several antibiotics and survived in feed; however, neither of the two isolates prevented S. parasitica infection when administered in feed.     

Synergistic effect of sodium selenite and Pediococcus acidilactici on growth,intestinal bacterial counts,selenium bioavailability,hepatic enzymes and non‐specific immune response in rainbow trout (Oncorhynchus mykiss)

Lactic acid bacteria (LAB) have a crucial role in inorganic selenium metabolism as well as their known desirable effects on fish. In this study, the synergistic effects of dietary sodium selenite and Pediococcus acidilactici on growth performance, intestinal bacterial counts, selenium bioavailability, hepatic antioxidant enzyme thioredoxin reductase activity and hepatic glycolytic enzyme activity that is hexokinase, phosphofructokinase and pyruvate kinase, and non‐specific immune response such as serum lysozyme and complements C₃, C₄ and ACH₅₀ activity in rainbow trout (Oncorhynchus mykiss) were investigated. Thus, a 3 × 3 factorial design experiment was conducted with nine purified diets including three levels of sodium selenite (0, 1 and 2 mg/kg) and three levels of P. acidilactici (0, 7 and 9 log CFU/g). After 8 weeks of feeding, weight gain and specific growth rate were increased by increasing dietary sodium selenite and P. acidilactici levels compared to control (p < .05), whereas feed conversion ratio and condition factor was decreased by increasing dietary sodium selenite and P. acidilactici amounts in comparison with control (p < .05). Survival rate was not significantly affected among the experimental treatments (p > .05). Total cultivable bacterial populations after 4 and 8 weeks of the feeding trial were not significantly different among the dietary treatments, while LAB levels were higher in P. acidilactici‐fed groups than in control and selenium‐fed groups (p < .05). Selenocysteine, methylselenocysteine and selenomethionine levels in the intestine of rainbow trout were increased by increasing the sodium selenite and P. acidilactici levels (p < .05), and selenocysteine was found the most selenium species in the trout intestine. The quantity of total selenium in the whole body, intestine, blood, liver and muscle of rainbow trout were increased by increasing the amounts of sodium selenite and P. acidilactici compared to control (p < .05). Hepatic thioredoxin reductase and hexokinase activity were increased by increasing dietary selenium and P. acidilactici levels in comparison to control (p < .05), whereas phosphofructokinase and pyruvate kinase activity in the liver of rainbow trout were not significantly different between the dietary treatments. Serum lysozyme, complements C₃, C₄ and ACH₅₀ activity were enhanced by increasing dietary selenium and P. acidilactici levels compared to control (p < .05). The most synergistic effects of dietary supplements on growth and metabolism of rainbow trout were obtained at 2 mg/kg sodium selenite and 7 log CFU/g P. acidilactici. The findings revealed the synergistic effect of dietary selenium and P. acidilactici on growth and metabolism in rainbow trout (O. mykiss).     

Timing to promote initial swimbladder inflation by surface film removal in Pacific bluefin tuna,Thunnus orientalis (Temminck and Schlegel), larvae

This study investigated the optimal timing of day to promote initial swimbladder inflation (ISI) for improved Pacific bluefin tuna (PBT), Thunnus orientalis, larval survival. Larval swimbladder inflation frequency was compared based on three experiments using different time schemes of surface film removal (SFR) from 3 to 9 days post hatch (dph). SFR was conducted from 05:00 to 19:00 hours (light period: S.5–19), 19:00 to 05:00 hours (dark period: S.19–5), 08:00 to 19:00 hours (S.8–19) and the entire day (S.24) in Experiment 1; from 08:00 to 19:00 hours (S.8–19‐E2), 08:00 to 13:00 hours (S.8–13), 13:00 to 19:00 hours (S.13–19) in Experiment 2; and from 13:00 to 16:00 hours (S.13–16), 16:00 to 19:00 hours (S.16–19), 18:00–19:00 hours (S.18–19) in Experiment 3. The swimbladder inflation frequency at the experiment termination (9 dph) was significantly higher (P < 0.001) in S.24 (91.1 ± 5.7%), S.5–19 (92.2 ± 5.1%) and S.8–19 (93.3 ± 3.4%) than in S.19–5 (11.1 ± 5.1%) in Experiment 1, and remarkably higher in S.8–19‐E2 (81.7%) and S.13–19 (88.3%) than in S.8–13 (0.0%) in Experiment 2, and significantly higher (P < 0.001) in S.16–19 (84.4 ± 5.1%) and S.18–19 (70.0 ± 12.0%) than in S.13–16 (7.8 ± 3.9%) in Experiment 3. These results suggest that the optimal timing to promote larval ISI by SFR is a few hours before the end of light period (16:00–19:00 hours) from 3 to 9 dph.     

Efficacy of a polyvalent injectable vaccine against Flavobacterium psychrophilum administered to rainbow trout (Oncorhynchus mykiss L.)

Flavobacterium psychrophilum is one of the most important pathogens affecting cultured rainbow trout (Oncorhynchus mykiss). Recent information from UK salmonid farms showed country‐wide distribution of genetically and serologically divergent clones, which has hampered the development of a vaccine for rainbow trout fry syndrome. The current study assessed the efficacy of an injectable polyvalent vaccine containing formalin‐inactivated F. psychrophilum in rainbow trout. The vaccine was formulated with an oil adjuvant (Montanide ISA 760VG) or formalin‐killed cells alone. Duplicate groups of trout (60 ± 13 g) were given phosphate‐buffered saline or vaccine formulated with Montanide by intra‐peritoneal (i.p.) injection and challenged by intra‐muscular (i.m.) injection with a homologous and a heterologous isolate of F. psychrophilum at 525 degree days post‐vaccination (dd pv). Significant protection was achieved in vaccinated fish (p = 0.0001, RPS 76% homologous, 88% heterologous). Efficacy of the adjuvanted vaccine was also demonstrated by heterologous challenge at 1155 dd pv resulting in 100% protection, whereas survival in the un‐adjuvanted group was not significantly different from control fish. Levels of specific antibody at 1155 dd pv, as measured by ELISA, were significantly higher in the fish vaccinated with adjuvant when compared with unvaccinated fish.     

Egg oxidation status,antioxidant enzyme activities,lipid classes,fatty acid composition profile and embryo survival rates during in vitro oocyte ageing in tench Tinca tinca (Linnaeus, 1758)

The effect of in vitro storage of oocytes on embryos survival rate, egg oxidation status, antioxidant enzyme activities, lipid classes and fatty acid composition profile was investigated in common tench Tinca tinca. In order to identify the role of oxidative stress in the progress of oocyte ageing the levels of thiobarbituric acid reactive substances (TBARS) and carbonyls as indicators of lipid and protein oxidation were measured and the activity of antioxidant enzymes were examined. Stripped ova from six females were stored in cell culture plates at 20°C for up to 10 hr post‐stripping (HPS). The stored ova were fertilized and the embryo survival rates were assessed. The results indicated that tench eggs could be successfully stored in vitro for 4 hr after stripping at 20°C. Superoxide dismutase enzyme activity increased at 6 HPS, whereas glutathione peroxidase and glutathione reductase activities decreased in oocytes during in vitro storage (p < .05) at 4 and 8 HPSs, respectively. The level of malondialdehyde did not show any significant changes during the progress of oocyte ageing. Carbonyls increased up to 2 HPS and thereafter decreased significantly. However, ova ageing did not affect the main lipid class composition and the fatty acid composition of the eggs. Lower quality eggs exhibited lower levels of cholesterol but higher levels of triacylglycerol.     

Probiotic characteristics and aflatoxin B1 binding ability of Debaryomyces hansenii and Kazaschtania exigua from rainbow trout environment

The aim of this study was to evaluate probiotic properties and the aflatoxin B₁ adsorption ability of yeasts isolated from rainbow trout intestine and fish feed to assess their use in the formulation of feed additives. Growth at pH 2, bacterial pathogens inhibition, bacterial pathogens co‐aggregation, autoaggregation, homologous and heterologous inhibition against lactic acid bacteria were evaluated. Moreover, aflatoxin B₁ adsorption was tested. All strains were able to maintain viable (10⁷ cells/ml) at pH 2. All strains isolated from intestine were identified as Kazaschtania exigua, while strains isolated from feed were all identified as Debaryomyces hansenii. Kazaschtania exigua RC035 and RC037 showed the strongest antimicrobial activity while K. exigua RC037 and RC038 were the most efficient co‐aggregating bacterial pathogens. All strains exhibited strong autoaggregation. None of the tested yeast strains showed homologous inhibition towards other yeasts and heterologous inhibition towards lactic acid bacteria strains. Debaryomyces hansenii RC031 demonstrated aflatoxin B₁ adsorption capacity (21%). The results of the present study indicate that select strains of Kazaschtania exigua and D. hansenii showed potential to improve the health of rainbow trout by inhibiting pathogens and binding AFB₁ and their use as probiotics may improve the production of rainbow trout in aquaculture systems.     

Association of a specific major histocompatibility complex class IIβ single nucleotide polymorphism with resistance to lactococcosis in rainbow trout,Oncorhynchus mykiss (Walbaum)

Major histocompatibility complex (MHC) loci encode glycoproteins that bind to foreign peptides and initiate immune responses through their interaction with T cells. MHC class II molecules are heterodimers consisting of α and β chains encoded by extremely variable genes; variation in exon 2 is responsible for the majority of observed polymorphisms, mostly concentrated in the codons specifying the peptide‐binding region. Lactococcus garvieae is the causative agent of lactococcosis, a warm‐water bacterial infection pathogenic for cultured freshwater and marine fish. It causes considerable economic losses, limiting the profitability and development of fish industries in general and the intensive production of rainbow trout, Oncorhynchus mykiss (Walbaum), in particular. The disease is currently controlled with vaccines and antibiotics; however, vaccines have short‐term efficacy, and increasing concerns regarding antibiotic residues have called for alternative strategies. To explore the involvement of the MHC class II β‐1 domain as a candidate gene for resistance to lactococcosis, we exposed 400 rainbow trout to naturally contaminated water. One single nucleotide polymorphism (SNP) and one haplotype were associated with resistance (P < 0.01). These results are promising for using MHC class IIβ as a molecular marker in breeding rainbow trout resistant to lactococcosis.     

Evaluation of dietary natural mineral materials as an antibiotic replacer on growth performance,non‐specific immune responses and disease resistance in rainbow trout,Oncorhynchus mykiss

We evaluated the effects of some dietary natural mineral materials as an antibiotic replacer based on growth performance, non‐specific immune responses and disease resistance in juvenile and subadult rainbow trout, Oncorhynchus mykiss. First experiment, juvenile rainbow trout averaging 2.7 ± 0.02 g (mean ± SD) were fed one of the six experimental diets; a basal commercial diet as a control (CON), CON with oxytetracycline (OTC), with yellow loess (YL), with Macsumsuk^® (MS), with Song‐Gang^® stone (SG) and with barley stone (BS) at 0.4% of each diet. At the end of 8‐week feeding trial, weight gain (WG), specific growth rate (SGR), feed efficiency (FE) and protein efficiency ratio (PER) of fish fed YL diet were significantly higher than those of fish fed CON diet. Non‐specific immune responses such as superoxide dismutase (SOD), myeloperoxidase (MPO), lysozyme (LYS) activity and oxidative radical production of fish fed YL diet were higher than those of fish fed CON diet. At the end of 15 days of challenge test with Aeromonas salmonicida, average cumulative survival rate of fish fed YL diet was significantly higher than that of fish fed BS and CON diets. However, there were no significant differences among fish fed YL, SG and OTC diets. Second experiment, subadult rainbow trout averaging 261.5 ± 3.5 g (mean ± SD) were fed one of the four experimental diets for 22 weeks: CON, and CON with OTC, YL or SG at 0.4% of each diet. At the end of feeding, growth performance of fish fed SG and YL diets was significantly higher than that of fish fed CON diet. Non‐specific immune responses in terms of SOD, MPO, LYS and NBT of fish fed SG and YL diets were significantly higher than those of fish fed CON diet. However, there were no significant differences among the fish fed YL, SG and OTC diets. The results indicate that dietary yellow loess or Song‐gang^® stone at 0.4% of diet could replace oxytetracycline in juvenile and subadult rainbow trout.